CN103285072A - Hyperoside-containing extract and medical application thereof - Google Patents
Hyperoside-containing extract and medical application thereof Download PDFInfo
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Abstract
The invention belongs to the field of traditional Chinese medicines and medicinal chemistry and relates to a hyperoside-containing extract and medical application thereof. The invention further relates to an extracting containing hyperoside and/or a medicinal salt thereof. Specifically, the invention relates to an applicaiton of hyperoside or the medical salt thereof or the extract disclosed by the invention or a pharmaceutical composition of the extract in preparation of a drug for treating and/or preventing and/or adjunctively treating obesity or obesity-related diseases or preparing a weight-reducing drug, which can be used for effectively treating and/or preventing and/or adjunctively treating obesity or obesity-related diseases or can be used for reducing weight.
Description
Technical field
The invention belongs to Chinese medicine and pharmaceutical chemistry field, relate to the extract and the medical usage thereof that comprise hyperin.
Background technology
Obesity has become worldwide epidemic diseases as a kind of general incretion metabolism disease.It not only influences figure and activity, and closely related with diseases such as hyperlipemia, atherosclerosis, coronary heart disease, diabetes.Because alimentary abstinence and increase motion can not reach satisfied fat-reducing effect usually, so Drug therapy has become the critical treatment means of obesity.
The fat reason that takes place is many-sided, and as inherited genetic factors, environmental factors, dietary habit etc., wherein high fat diet is the major reason that causes fat.In recent years, Drug therapy is one of effective means for the treatment of of obesity, reaches therapeutic purposes by reducing fat absorption.Pancreas and gastric lipase are that fat digestion absorbs necessary in the intestinal.Fat in the food is absorbed at intestinal after being hydrolyzed to monoacylglycerol and free fatty, and synthctic fat again in vivo causes athero then, finally can cause fat.Use lipase inhibitor and can suppress effectively that lipase reaches the purpose that reduces fat absorption, control and treatment obesity to the decomposition catalytic action of fat in the intestinal.Thereby the effective lipase inhibitor of development and application, be subjected to people's attention.
Though novel appetrol action target spot constantly is found, also to develop some and it seems promising medicine, most medicines all also are in the early development stage, and its curative effect and safety are on the knees of the gods.At present, U.S. FDA has only been ratified 2 kinds of medicines and can be used for bariatrician: sibutramine and orlistat for a long time.But these two kinds of medicines have the side effect of tangible untoward reaction: sibutramine to mainly contain thirsty, constipation, dizziness and insomnia, the untoward reaction of orlistat mainly is gastrointestinal symptom, common have diarrhoea stomachache, oiliness speckle, a flatulence etc., and its long-term effect also awaits further evaluation.
Hyperin belongs to the flavonol glycosides compounds, Latin is called Hyperoside, English Hyperin by name, and different name is VincetoxicosideB, hyperin, chemistry is by name, and Quercetin-(Quercetin-3-O-β-galactoside), structure is shown in following formula I for 3-O-beta galactose glycosides.
Formula I
Hyperin is present in the various plant bodies, for example, and fruit and the herb of Hypericaceae, Rosaceae, campanulaceae, lip type section, little Pi section, Ericaceae, kurrajong etc.The bibliographical information hyperin has antiallergic, spasmolytic, diuresis, significantly cough-relieving, blood pressure lowering, cholesterol reducing and blood fat, protein assimilation, gastric mucosa and hepatic tissue protection, effect such as enhance immunity and antidepressant, but study at present maximum be local and the central analgesia effect reach protective effect to cardiovascular and cerebrovascular vessel (Li Minfang etc. the hyperin Advance on Pharmacological Activities. Chinese Chinese medicine information magazine, 2008,15 (4): 102-104; Zhang Chengwei etc. the hyperin Advance on Pharmacological Activities. Anhui medicine, 2007,11 (11): 961-962).
At present, still need and to find the new medicine of obesity that is used for preventing and treating.
Summary of the invention
The inventor is through in depth exploring and analyzing, carried out a large amount of research, (Latin is called Aeschynomene indica L, annual suffruticose draft from a kind of plant Herba Aeschynonenes Indicae of pulse family, be born in flush or the waterside, be distributed in ground such as North China, East China, Central-South, southwest.Different name: Herba Aeschynonenes Indicae, the water antler, close bright grass, the raft bean, multiple different names such as wild Fructus Gleditsia) separate in and obtained hyperin, and be surprised to find, this chemical compound has good fat-reducing effect, has as treating and/or preventing and/or the potentiality of auxiliary treatment obesity or obesity relevant disease medicine or slimming medicine.Following invention is provided thus:
One aspect of the present invention relates to the extract that comprises hyperin and/or its officinal salt.Particularly, described extract can be ethanol extraction.
Each described extract according to the present invention, wherein, the content of hyperin is more than or equal to 0.73% (w/w) in the described extract; Preferably, for more than or equal to 3.28% (w/w); More preferably, for more than or equal to 8.20% (w/w); Further preferably, for more than or equal to 32.8% (w/w).
Each described extract according to the present invention, wherein, described extract is the leguminous plant extract; Particularly, be the complexing resistance spp plant extract; More specifically, be the Herba Aeschynonenes Indicae extract.
Each described extract according to the present invention, it is as follows 1 years old), step 1) to 2) or step 1) to 3) make:
1) gets leguminous plant or complexing resistance spp plant or Herba Aeschynonenes Indicae aerial parts 1kg, add 6-10 times of volume 30%-95% ethanol (v/v) (preferred 50%-70% ethanol, for example 50%, 60%, 70%) reflux, extract, 1 time or repeatedly (preferred 3 times) are at every turn more than or equal to 1 hour (preferred 1-2 hour);
2) sucking filtration, filtrate merge, and the 1.8-2.2 that is concentrated into crude drug weight doubly (for example 1.8,1.9,2.0,2.1 or 2.2 times, preferred 2 times), centrifugal, get supernatant;
3) supernatant is injected macroporous adsorptive resins (preferred D101 macroporous resin), after removing impurity with the alcoholic solution eluting that is less than or equal to 10%, reuse is more than or equal to 11% ethanol (be preferably greater than or equal 45%) eluting (for example 2 times of column volumes), collects lyophilizing after ethanol more than or equal to 11% (be preferably greater than or equal 45%) the eluting partial concentration.
Each described extract according to the present invention, wherein, the preparation of described extract also comprises the steps:
4) product that obtains in the step 3) is mixed sample with an amount of (can dissolve the amount of described product fully) dichloromethane and methanol mixed liquor dissolving in 1: 1 back with silica gel (about 50g), carry out silica gel column chromatography (500g silica gel dress post, the 200-300 order), ethyl acetate: methanol: water=10: 2: 1 eluting; Collect first column volume (flow point TZJ-1), concentrate and lyophilizing;
Preferably, also comprise the steps:
5) product that obtains in the step 4) is carried out sephadex LH-20 gel filtration chromatography and separate, detect according to TLC and collect and concentrate.
Another aspect of the present invention relates to a kind of pharmaceutical composition, and it comprises each described extract among the present invention of effective dose, and/or comprises hyperin and/or its officinal salt; Alternatively, described pharmaceutical composition also comprises pharmaceutically acceptable carrier or adjuvant.
Usually pharmaceutical composition of the present invention contains hyperin and/or its officinal salt of 0.1-90 weight % and/or comprises hyperin and/or the extract of its officinal salt.Pharmaceutical composition can prepare according to methods known in the art.When being used for this purpose, if desired, hyperin and/or its officinal salt and one or more solids or liquid medicine excipient and/or adjuvant can be combined, make the suitable administration form or the dosage form that can be used as human.
Hyperin and/or its officinal salt or contain its pharmaceutical composition can the unit dosage form administration, route of administration can be intestinal or non-intestinal, as oral, muscle, subcutaneous, nasal cavity, oral mucosa, skin, peritoneum or rectum etc.Form of administration is tablet, capsule, drop pill, aerosol, pill, powder, solution, suspensoid, Emulsion, granule, liposome, transdermal agent, buccal tablet, suppository, lyophilized injectable powder etc. for example.Can be ordinary preparation, slow releasing preparation, controlled release preparation and various particulate delivery system.For the unit form of administration is made tablet, can be extensive use of various carrier well known in the art.Example about carrier is, for example diluent and absorbent are as starch, dextrin, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, glucose, carbamide, calcium carbonate, kaolin, microcrystalline Cellulose, aluminium silicate etc.; Wetting agent and binding agent are as water, glycerol, Polyethylene Glycol, ethanol, propanol, starch slurry, dextrin, syrup, Mel, glucose solution, mucialga of arabic gummy, gelatine size, sodium carboxymethyl cellulose, lac, methylcellulose, potassium phosphate, polyvinylpyrrolidone etc.; Disintegrating agent, for example dry starch, alginate, agar powder, laminaran, sodium bicarbonate and citric acid, calcium carbonate, polyoxyethylene, Sorbitol fatty acid ester, dodecyl sodium sulfate, methylcellulose, ethyl cellulose etc.; Disintegrate inhibitor, for example sucrose, glyceryl tristearate, cocoa butter, hydrogenation wet goods; Absorption enhancer, for example quaternary ammonium salt, sodium lauryl sulphate etc.; Lubricant, for example Pulvis Talci, silicon dioxide, corn starch, stearate, boric acid, liquid paraffin, Polyethylene Glycol etc.Tablet further can also be made coated tablet, for example sugar coated tablet, thin membrane coated tablet, ECT, or double-layer tablet and multilayer tablet.For pill is made in the administration unit, can be extensive use of various carrier well known in the art.Example about carrier is, for example diluent and absorbent are as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, polyvinylpyrrolidone, Ge luc i r e, Kaolin, Pulvis Talci etc.; Binding agent such as arabic gum, Tragacanth, gelatin, ethanol, Mel, liquid sugar, rice paste or batter etc.; Disintegrating agent is as agar powder, dry starch, alginate, dodecyl sodium sulfate, methylcellulose, ethyl cellulose etc.For suppository is made in the administration unit, can be extensive use of various carrier well known in the art.Example about carrier is, for example the ester of Polyethylene Glycol, lecithin, cocoa butter, higher alcohol, higher alcohol, gelatin, semi-synthetic glyceride etc.For capsule is made in the administration unit, effective ingredient hyperin and/or its officinal salt are mixed with above-mentioned various carriers, and the mixture that will obtain thus places hard obviously capsule or soft capsule.Also effective ingredient hyperin and/or its officinal salt can be made microcapsule, be suspended in and form suspensoid in the aqueous medium, in the hard capsule of also can packing into or make injection and use.For injection preparation is made in the administration unit, as solution, Emulsion, lyophilized injectable powder and suspensoid, can use this area all diluent commonly used, for example, water, ethanol, Polyethylene Glycol, 1, the isooctadecanol of ammediol, ethoxylation, the isooctadecanol of polyoxyization, Polyoxyethylene Sorbitol Fatty Acid Esters etc.In addition, to ooze injection in order preparing etc., can in injection preparation, to add proper amount of sodium chloride, glucose or glycerol, in addition, can also add conventional cosolvent, buffer agent, pH regulator agent etc.
In addition, as needs, also can in pharmaceutical preparation, add coloring agent, antiseptic, spice, correctives, sweeting agent or other material.
The dosage of hyperin and/or its officinal salt or pharmaceutical composition of the present invention depends on many factors, for example to prevent or treat character and the order of severity of disease, the sex of patient or animal, age, body weight and individual reaction, used particular compound, route of administration and administration number of times etc.Above-mentioned dosage can the single dose form or be divided into several, for example two, three or four dosage form administrations.
Term used herein " compositions " means and comprises the product of respectively specifying composition that comprises specified amount, and directly or indirectly from any product of the combination results of respectively specifying composition of specified amount.
The reactive compound amount of gained can change the actual dose level of each active component in the pharmaceutical composition of the present invention, so that can effectively obtain required therapeutic response at concrete patient, compositions and administering mode.The dosage level fibrous root is selected according to activity, route of administration, the order of severity of the patient's condition for the treatment of and the patient's to be treated patient's condition and the medical history of particular compound.But the way of this area is that the dosage of chemical compound increases dosage gradually from being lower than for obtaining the level that required therapeutic effect requires, up to obtaining required effect.
Of the present inventionly relate in one aspect among hyperin or its officinal salt or the present invention each described extract again or each described pharmaceutical composition of the present invention treats and/or prevents and/or the medicine of auxiliary treatment obesity or obesity relevant disease or the purposes in the slimming medicine in preparation; Particularly, described obesity relevant disease is hyperlipemia, atherosclerosis, coronary heart disease or diabetes.
Of the present inventionly relate in one aspect to a kind for the treatment of and/or preventing and/or auxiliary treatment obesity or obesity relevant disease or ways of preventing obesity again, comprise the step of each described extract among the hyperin that gives experimenter's effective dose or its officinal salt or the present invention or each described pharmaceutical composition of the present invention.
When being used for above-mentioned treat and/or prevent or during auxiliary treatment, the hyperin and/or its officinal salt that treat and/or prevent effective dose can be used with pure form, perhaps use with the acceptable ester of pharmacy or prodrug forms (under the situation that has these forms).Perhaps, can accept the pharmaceutical composition administration of excipient to contain hyperin and/or its officinal salt and one or more medicines.But the total consumption per day that it should be understood that hyperin and/or its officinal salt or pharmaceutical composition of the present invention must be examined the doctor by the master and maked decision in the medical judgment scope reliably.For any concrete patient, the concrete horizontal fibrous root for the treatment of effective dose is decided according to multiple factor, and described factor comprises the order of severity of the obstacle for the treatment of and this obstacle; The activity of the particular compound that adopts; The concrete compositions that adopts; Patient's age, body weight, general health situation, sex and diet; The administration time of the particular compound that adopts, route of administration and excretion rate; The treatment persistent period; The medicine that is used in combination or uses simultaneously with the particular compound that adopts; And the known similar factor of medical field.For example, the way of this area is that the dosage of chemical compound increases dosage gradually from being lower than for obtaining the level that required therapeutic effect requires, up to obtaining required effect.In general, particularly people's dosage can be between the 0.001-1000mg/kg body weight/day, for example between the 0.01-100mg/kg body weight/day, for example between the 0.01-10mg/kg body weight/day for mammal for hyperin and/or its officinal salt.
Hyperin and/or its officinal salt or pharmaceutical composition of the present invention can effectively prevent and/or treatment and/or auxiliary treatment various diseases of the present invention or disease.
Of the present inventionly relate in one aspect to each described extract among hyperin or its officinal salt or the present invention again or each described pharmaceutical composition of the present invention suppresses lipase active or regulates the medicine of lipase active or the purposes in the reagent in preparation.
The method that relates in one aspect in vivo a kind of or vitro inhibition again or regulate lipase active of the present invention comprises the step of each described extract among the silk peach glycosides that uses effective dose or its officinal salt or the present invention or each described pharmaceutical composition of the present invention.
Of the present inventionly relate in one aspect to a kind of method for preparing hyperin or comprise the extract of hyperin again, it is characterized in that using Herba Aeschynonenes Indicae to be raw material;
Particularly, comprise the steps (for example Xia Mian step 1), step 1) to 2), step 1) to 3), step 1) to 4), step 1) to 5) or step 1) to 6)):
1) gets leguminous plant or complexing resistance spp plant or Herba Aeschynonenes Indicae aerial parts 1kg, add 6-10 times of volume 30%-95% ethanol (v/v) (preferred 50%-70% ethanol, for example 50%, 60%, 70%) reflux, extract, 1 time or repeatedly (preferred 3 times) are at every turn more than or equal to 1 hour (preferred 1-2 hour);
2) sucking filtration, filtrate merge, and the 1.8-2.2 that is concentrated into crude drug weight doubly (for example 1.8,1.9,2.0,2.1 or 2.2 times, preferred 2 times), centrifugal, get supernatant;
3) supernatant is injected macroporous adsorptive resins (preferred D101 macroporous resin), after removing impurity with the alcoholic solution eluting that is less than or equal to 10%, reuse is more than or equal to 11% ethanol (be preferably greater than or equal 45%) eluting (for example 2 times of column volumes), collects lyophilizing after ethanol more than or equal to 11% (be preferably greater than or equal 45%) the eluting partial concentration;
4) product that obtains in the step 3) is mixed sample with an amount of (can dissolve the amount of described product fully) dichloromethane and methanol mixed liquor dissolving in 1: 1 back with silica gel (about 50g), carry out silica gel column chromatography (500g silica gel dress post, the 200-300 order), ethyl acetate: methanol: water=10: 2: 1 eluting; Collecting first column volume (flow point TZJ-1) concentrates and lyophilizing;
5) product that obtains in the step 4) is carried out sephadex LH-20 gel filtration chromatography and separate, detect according to TLC and collect and concentrate;
6) product that obtains in the step 5) is carried out purification by half preparative high-performance liquid chromatographic;
Particularly,
The condition of half preparative high-performance liquid chromatographic is as follows described in the step 6):
Mobile phase A is acetonitrile: methanol=1: 1;
Mobile phase B is 0.2% acetic acid aqueous solution;
A∶B＝31∶69；
Flow velocity: 4mL/min;
Chromatographic column: C18 semi-preparative column (10*250mm, 5 μ m);
Retention time Rt is 10.3min;
It is 254 or 365nm that the DAD detector detects wavelength.
Leguminous plant or complexing resistance spp plant or the Herba Aeschynonenes Indicae purposes in preparation hyperin or its officinal salt or each described extract of the present invention that relates in one aspect to again of the present invention.
Among the present invention,
Term " effective dose " refers to realize treating, prevent, alleviate and/or alleviating the dosage of disease of the present invention or disease in the experimenter.
Term " experimenter " can refer to that patient or other accept among hyperin or its officinal salt or the present invention each described extract or each described pharmaceutical composition of the present invention to treat, to prevent, to alleviate and/or to alleviate the animal of disease of the present invention or disease, particularly mammal, for example people, Canis familiaris L., monkey, cattle, horse etc.
Term " disease and/or disease " refers to a kind of condition of described experimenter, and this condition is relevant with disease of the present invention and/or disease.
In the present invention, if not otherwise specified, the concentration of described methanol solution (aqueous solution) or alcoholic solution (aqueous solution) all refers to volumetric concentration (v/v).
Herba Aeschynonenes Indicae described in the present invention if not otherwise specified, preferably, is the aerial parts of Herba Aeschynonenes Indicae.
The beneficial effect of the invention
Hyperin or its officinal salt or extract of the present invention or pharmaceutical composition of the present invention can suppress lipase active effectively, reduce fat absorption, suppress weight increase, can be used in and treat and/or prevent and/or auxiliary treatment obesity or obesity relevant disease or fat-reducing.
Description of drawings
Fig. 1: last figure is the collection of illustrative plates that detects under the 254nm wavelength, and figure below is the collection of illustrative plates that detects under the 365nm wavelength.
The specific embodiment
Be described in detail below in conjunction with the embodiment of the present invention of embodiment, but it will be understood to those of skill in the art that the following example only is used for explanation the present invention, and should not be considered as limiting scope of the present invention.Unreceipted actual conditions person among the embodiment carries out according to the condition of normal condition or manufacturer's suggestion.The unreceipted person of production firm of agents useful for same or instrument, being can be by the conventional products of commercial acquisition.
Embodiment 1: the separation and purification of hyperin and activity rating (hyperin can be purchased)
Get Herba Aeschynonenes Indicae (available from the positive medical company limited of Nanyang health) aerial parts 1kg, add 6-10 times of volume (the 1st time be 8 times of volumes, be for 2 times 6 times of volumes, be for 3 times 6 times of volumes) 50% ethanol (v/v) reflux, extract, 3 times, each 1 hour; Sucking filtration, filtrate merges, and obtains amalgamation liquid, concentrates amalgamation liquid to 2 times of crude drug weight, and is centrifugal, gets supernatant; Supernatant is injected the D101 macroporous resin, and after 10% alcoholic solution (v/v) eluting was removed impurity, 2 times of column volumes of reuse 45% ethanol (v/v) eluting were collected lyophilizing after 45% ethanol (v/v) the eluting partial concentration, namely get Herba Aeschynonenes Indicae effective site 50g.
Herba Aeschynonenes Indicae effective site is dissolved back 50g left and right sides silica gel mixed sample with an amount of (can dissolve the amount of Herba Aeschynonenes Indicae effective site fully) dichloromethane and 1: 1 mixed liquor of methanol, 500g silica gel dress post (200-300 order), carry out silica gel column chromatography, ethyl acetate: methanol: water=10: 2: 1 eluting.Collect first column volume (flow point TZJ-1) and concentrate and lyophilizing, get dry powder 20g.
Get TZJ-1 dry powder 20g, carrying out sephadex LH-20 gel filtration chromatography separates, detect according to TLC that (the TLC testing conditions is the purification on normal-phase silica gel plate: developing solvent is dichloromethane-ethyl acetate-methanol-water=1: 5: 1: 0.3, the Rf value of hyperin is about 0.36,), collect and concentrate the position TZJ-1E (2.5g) that contains hyperin.Carry out purification with half preparative high-performance liquid chromatographic at last and obtain the about 250mg of hyperin.
Liquid-phase condition: mobile phase A is acetonitrile: methanol=1: 1, B are 0.2% acetic acid aqueous solution
A: B=31: 69; Flow velocity: 4mL/min; Chromatographic column: C18 semi-preparative column (10*250mm, 5 μ m); Retention time Rt is 10.3min; It is 254 or 365nm (Fig. 1) that the DAD detector detects wavelength.
In addition, after testing, above the content of hyperin in the extract that obtains of each step as follows:
Content in amalgamation liquid is 0.73%, and the content of the effective site that the process macroporous resin obtains is 3.28%, is 8.20% through the content in the products therefrom behind the silica gel column chromatography, and the content of process sephadex LH-20 gel section is 32.8%.
Embodiment 2: (the body fat enzyme suppresses to live hyperin to the mice fat absorption and effect
The detection of property)
Fatty tissue mainly is made up of the adipose cell that is rich in triglyceride, and triglyceride is the main storage form of body fat mass in human body, is one of important indicator of lipid metabolism.Lipase is a kind of special ester linkage hydrolyzing enzyme, and it can act on the ester bond of triglyceride, makes triglyceride be degraded to diglyceride, monoglyceride, glycerol and fatty acid.Thereby can infer that the activity of this triglyceride concentration and lipase and the amount of fat absorption have direct relation.
Concrete experimental technique can reference: Sugiyama H, Akazome Y, Shoji T, etal.Oligomeric procyanidins in apple polyphenol are main activecomponents for inhibition of pancreatic lipase and triglycerideabsorption.J Agric Food Chem.2007,55:4604-4609, also can be with reference to following step:
1. experiment material
Animal: male ICR mouse is divided into matched group and administration group (by body weight principle of reciprocity random packet); About body weight-30g; Animal origin-purchase in Beijing Vital River Experimental Animals Technology Co., Ltd.
Hyperin: according to the preparation of the method among the embodiment 1, also can be purchased;
Positive control medicine orlistat (Orlistat) is purchased in gold as big pharmacy;
Anticoagulant EDTA purchases in Beijing chemical reagents corporation;
Semen Maydis oil is purchased in the supermarket and is sent out the supermarket;
Measure the content of triglyceride test kit, purchase in Zhongsheng Beikong Biological Science ﹠ Technology Co., Ltd..
2. experimental technique
A. grouping, administration
Male ICR mouse about 30g is divided into matched group and administration group, 8 every group.The back administration is can't help about water 20h in fasting, and matched group is irritated stomach with the dosage of 5mL/kg and given Semen Maydis oil, when the administration group then gives isodose Semen Maydis oil, gives the reagent that is subjected to of various dose, and wherein the dosage of positive drug orlistat is 12.5mg/kg.2h cuts tail and gets blood after giving Semen Maydis oil, preparation blood plasma (EDTA anticoagulant).With the middle north control kit measurement content of triglyceride of giving birth to, calculate blood fat rising suppression ratio at last.
B. observation index and assay method
Detect index: plasma triglyceride concentration
Assay method: with reference to the survey triglyceride test kit description of Zhongsheng Beikong Biological Science ﹠ Technology Co., Ltd.
3. experimental result
Shown in following table 1.
Table 1: mice fat meal stress test
By table 1 as seen, hyperin has anti-fat absorption effect, can suppress lipase active (for example suppressing lipase active in vivo) effectively.
Embodiment 3: hyperin is to the influence of auxotype obesity mice body weight
Experimental technique can reference: the Liu quintessence, and Zhang Yong, Yu Xiaoming, etc. medium-chain fatty acid improves the effect of the lipoprotein levels of high lipid food short-term and the long-term C57BL/6J of nursing mice. experiment preventive medicine, 2011,18 (9): 1610-1613; Li Tao, nationality is kept tie, Zhou Feng, etc. red tea fungus is to the research of diet induced obesity mice body weight control. Food Science, 2009,30 (11): 246-251; Also can be with reference to following step:
1. experiment material
Animal: C57BL/6J male mice (3-4 age in week) is divided into matched group and administration group (by body weight principle of reciprocity random packet); About body weight-16-18g; Animal origin-available from Beijing Vital River Experimental Animals Technology Co., Ltd.;
High lipid food D12492 and low fat thermal energy fodder D12450B are available from the farsighted peaceful thing in Shanghai Science and Technology Ltd.;
Hyperin: according to the preparation of the method among the embodiment 1, also can be purchased;
Positive control medicine orlistat (Orlistat), available from gold as big pharmacy.
2. experimental technique
A. modeling, grouping and administration
Feed normal diet 7d with the C57BL/6J male mice under experimental situation, weigh, be divided into 2 groups at random: to finishing to use the low fat forage feed, all the other mices all feed high lipid food to blank group mice (totally 10) from the experiment beginning.After high lipid food fed for 4 weeks, the body weight of mice surpassed 20% conduct experiment obesity mice of low fat forage feed mice body weight.The experiment obesity mice is divided into 4 groups, model contrast, positive control (orlistat of 25mg/kg), hyperin high dose group (100mg/kg), and hyperin low dose group (50mg/kg), is irritated stomach every day 1 time, the back end experiment of 7 week of successive administration by 10 every group.
B. observation index
Body weight
3. experimental result
Shown in following table 2.
Table 2: hyperin is to the influence of auxotype obesity mice body weight
* p＜0.05 is compared with model control group
By table 2 as seen, initial body weight difference not statistically significant (p＞0.05) is respectively organized in experiment; When treatment finished, model control group and normal control group compared, and body weight significantly raises; Positive controls, hyperin high dose group, hyperin low dose group and model control group compare, and body weight significantly reduces, and show that hyperin has the effect of the mice of inhibition weight increase, can be used in control and/or auxiliary treatment obesity or its relevant disease.
Although the specific embodiment of the present invention has obtained detailed description, it will be understood to those of skill in the art that.According to disclosed all instructions, can carry out various modifications and replacement to those details, these change all within protection scope of the present invention.Four corner of the present invention is provided by claims and any equivalent thereof.
Claims (10)
1. the extract that comprises hyperin and/or its officinal salt.
2. extract according to claim 1, wherein, the content of hyperin is more than or equal to 0.73% (w/w) in the described extract; Preferably, for more than or equal to 3.28% (w/w); More preferably, for more than or equal to 8.20% (w/w); Further preferably, for more than or equal to 32.8% (w/w).
3. extract according to claim 1 and 2, wherein, described extract is the leguminous plant extract; Particularly, be the complexing resistance spp plant extract; More specifically, be the Herba Aeschynonenes Indicae extract.
4. extract according to claim 3, it is by following step 1), step 1) to 2), step 1) to 3), step 1) to 4) or step 1) to 5) make:
1) gets leguminous plant or complexing resistance spp plant or Herba Aeschynonenes Indicae aerial parts 1kg, add 6-10 times of volume 30%-95% ethanol (v/v) reflux, extract, 1 time or repeatedly, at every turn more than or equal to 1 hour;
2) sucking filtration, filtrate merges, and is concentrated into the 1.8-2.2 of crude drug weight doubly, and is centrifugal, gets supernatant;
3) supernatant is injected macroporous adsorptive resins, with after being less than or equal to 10% alcoholic solution eluting and removing impurity, reuse is collected more than or equal to lyophilizing after 11% the ethanol elution partial concentration more than or equal to 11% ethanol elution;
4) product that obtains in the step 3) is used silica gel mixed sample with an amount of dichloromethane and methanol mixed liquor dissolving in 1: 1 back, carry out silica gel column chromatography, ethyl acetate: methanol: water=10: 2: 1 eluting; Collect first column volume, concentrate and lyophilizing;
5) product that obtains in the step 4) is carried out sephadex LH-20 gel filtration chromatography and separate, detect according to TLC and collect and concentrate.
5. pharmaceutical composition, it comprises each described extract in the claim 1 to 4 of effective dose, and/or comprises hyperin and/or its officinal salt; Alternatively, described pharmaceutical composition also comprises pharmaceutically acceptable carrier or adjuvant.
6. each described extract or the described pharmaceutical composition of claim 5 treat and/or prevent and/or the medicine of auxiliary treatment obesity or obesity relevant disease or the purposes in the slimming medicine in preparation in hyperin or its officinal salt or the claim 1 to 4; Particularly, described obesity relevant disease is hyperlipemia, atherosclerosis, coronary heart disease or diabetes.
7. each described extract or the described pharmaceutical composition of claim 5 suppress lipase active or regulate the medicine of lipase active or the purposes in the reagent in preparation in hyperin or its officinal salt or the claim 1 to 4.
One kind in vivo or vitro inhibition or the method for regulating lipase active, comprise the step of each described extract in the silk peach glycosides that uses effective dose or its officinal salt or the claim 1 to 4 or the described pharmaceutical composition of claim 5.
9. a method for preparing hyperin or comprise the extract of hyperin is characterized in that using leguminous plant or complexing resistance spp plant or Herba Aeschynonenes Indicae to be raw material;
Particularly, comprise the steps:
1) gets leguminous plant or complexing resistance spp plant or Herba Aeschynonenes Indicae aerial parts 1kg, add 6-10 times of volume 30%-95% ethanol (v/v) reflux, extract, 1 time or repeatedly, at every turn more than or equal to 1 hour;
2) sucking filtration, filtrate merges, and is concentrated into the 1.8-2.2 of crude drug weight doubly, and is centrifugal, gets supernatant;
3) supernatant is injected macroporous adsorptive resins, with after being less than or equal to 10% alcoholic solution eluting and removing impurity, reuse is collected more than or equal to lyophilizing after 11% the ethanol elution partial concentration more than or equal to 11% ethanol elution;
4) product that obtains in the step 3) is used silica gel mixed sample with an amount of dichloromethane and methanol mixed liquor dissolving in 1: 1 back, carry out silica gel column chromatography, ethyl acetate: methanol: water=10: 2: 1 eluting; Collecting first column volume concentrates and lyophilizing;
5) product that obtains in the step 4) is carried out sephadex LH-20 gel filtration chromatography and separate, detect according to TLC and collect and concentrate;
6) product that obtains in the step 5) is carried out purification by half preparative high-performance liquid chromatographic;
Particularly,
The condition of half preparative high-performance liquid chromatographic is as follows described in the step 6):
Mobile phase A is acetonitrile: methanol=1: 1;
Mobile phase B is 0.2% acetic acid aqueous solution;
A∶B＝31∶69；
Flow velocity: 4mL/min;
Chromatographic column: C18 semi-preparative column (10*250mm, 5 μ m);
Retention time Rt is 10.3min;
It is 254 or 365nm that the DAD detector detects wavelength.
10. leguminous plant or complexing resistance spp plant or the Herba Aeschynonenes Indicae purposes in each described extract in preparation hyperin or its officinal salt or claim 1 to 4.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106306956A (en) * | 2016-08-24 | 2017-01-11 | 开平市水口镇卡摩商行 | Health-care drink |
| CN111150741A (en) * | 2020-01-22 | 2020-05-15 | 辽宁中医药大学 | Medical application of hyperin in relieving human aortic endothelial cell injury |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1308492A (en) * | 1998-07-09 | 2001-08-15 | Cv技术公司 | Hypericin and i(hypericum) extract: specific T-type calcium channel blocker, and their use as T-type calcium channel targeted therapeutics |
| CN1634325A (en) * | 2004-11-18 | 2005-07-06 | 李青山 | Apocynum extract and extracting method thereof |
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2012
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1308492A (en) * | 1998-07-09 | 2001-08-15 | Cv技术公司 | Hypericin and i(hypericum) extract: specific T-type calcium channel blocker, and their use as T-type calcium channel targeted therapeutics |
| CN1634325A (en) * | 2004-11-18 | 2005-07-06 | 李青山 | Apocynum extract and extracting method thereof |
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| 陈家源等: "田皂角中的脂溶性成分分析", 《广西科学》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106306956A (en) * | 2016-08-24 | 2017-01-11 | 开平市水口镇卡摩商行 | Health-care drink |
| CN111150741A (en) * | 2020-01-22 | 2020-05-15 | 辽宁中医药大学 | Medical application of hyperin in relieving human aortic endothelial cell injury |
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| CN103285072B (en) | 2015-09-09 |
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